中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2012年
7期
543-547
,共5页
胰腺肿瘤%甲基化%基因表达调控%5-杂氮脱氧胞嘧啶核苷
胰腺腫瘤%甲基化%基因錶達調控%5-雜氮脫氧胞嘧啶覈苷
이선종류%갑기화%기인표체조공%5-잡담탈양포밀정핵감
Pancreatic neoplasms%Methylation%Gene expression regulation%5-Aza-CdR
目的 研究5-杂氮脱氧胞嘧啶核苷(5-Aza-CdR)对胰腺癌细胞株Capan-2原钙黏蛋白8(protocadherin 8,PCDH8)基因DNA启动子甲基化的转录调控,以及联合吉西他滨对癌细胞生长抑制与凋亡的影响.方法 采用MTT法、流式细胞术检测5-Aza-CdR及联合吉西他滨对Capan-2细胞的抑制率与凋亡率.应用甲基化特异性PCR(MSP)、RT-PCR和Western blot的方法检测不同浓度5-Aza-CdR处理细胞前后PCDH8基因的甲基化状态、mRNA表达水平和蛋白表达水平.结果 5-Aza-CdR能使胰腺癌细胞Capan-2增长速度减慢,呈浓度依赖性,联合吉西他滨显著抑制细胞生长；5-Aza-CdR联合吉西他滨与单药组、对照组相比,可显著诱导细胞凋亡.不同浓度的5-Aza-CdR可逆转PCDH8基因在胰腺癌细胞Capan-2的甲基化,恢复mRNA表达水平和蛋白表达水平,并呈一定的浓度正相关.结论 5-Aza-CdR可有效逆转PCDH8基因在胰腺癌细胞株Capan-2的高甲基化状态,解除DNA高甲基化导致的基因沉默,重新诱导基因mRNA转录和蛋白表达,同时可抑制胰腺癌细胞生长,与吉西他滨有协同抗肿瘤作用.
目的 研究5-雜氮脫氧胞嘧啶覈苷(5-Aza-CdR)對胰腺癌細胞株Capan-2原鈣黏蛋白8(protocadherin 8,PCDH8)基因DNA啟動子甲基化的轉錄調控,以及聯閤吉西他濱對癌細胞生長抑製與凋亡的影響.方法 採用MTT法、流式細胞術檢測5-Aza-CdR及聯閤吉西他濱對Capan-2細胞的抑製率與凋亡率.應用甲基化特異性PCR(MSP)、RT-PCR和Western blot的方法檢測不同濃度5-Aza-CdR處理細胞前後PCDH8基因的甲基化狀態、mRNA錶達水平和蛋白錶達水平.結果 5-Aza-CdR能使胰腺癌細胞Capan-2增長速度減慢,呈濃度依賴性,聯閤吉西他濱顯著抑製細胞生長；5-Aza-CdR聯閤吉西他濱與單藥組、對照組相比,可顯著誘導細胞凋亡.不同濃度的5-Aza-CdR可逆轉PCDH8基因在胰腺癌細胞Capan-2的甲基化,恢複mRNA錶達水平和蛋白錶達水平,併呈一定的濃度正相關.結論 5-Aza-CdR可有效逆轉PCDH8基因在胰腺癌細胞株Capan-2的高甲基化狀態,解除DNA高甲基化導緻的基因沉默,重新誘導基因mRNA轉錄和蛋白錶達,同時可抑製胰腺癌細胞生長,與吉西他濱有協同抗腫瘤作用.
목적 연구5-잡담탈양포밀정핵감(5-Aza-CdR)대이선암세포주Capan-2원개점단백8(protocadherin 8,PCDH8)기인DNA계동자갑기화적전록조공,이급연합길서타빈대암세포생장억제여조망적영향.방법 채용MTT법、류식세포술검측5-Aza-CdR급연합길서타빈대Capan-2세포적억제솔여조망솔.응용갑기화특이성PCR(MSP)、RT-PCR화Western blot적방법검측불동농도5-Aza-CdR처리세포전후PCDH8기인적갑기화상태、mRNA표체수평화단백표체수평.결과 5-Aza-CdR능사이선암세포Capan-2증장속도감만,정농도의뢰성,연합길서타빈현저억제세포생장；5-Aza-CdR연합길서타빈여단약조、대조조상비,가현저유도세포조망.불동농도적5-Aza-CdR가역전PCDH8기인재이선암세포Capan-2적갑기화,회복mRNA표체수평화단백표체수평,병정일정적농도정상관.결론 5-Aza-CdR가유효역전PCDH8기인재이선암세포주Capan-2적고갑기화상태,해제DNA고갑기화도치적기인침묵,중신유도기인mRNA전록화단백표체,동시가억제이선암세포생장,여길서타빈유협동항종류작용.
Objective To investigate the effects of 5-Aza-CdR on the transcriptional regulation through methylation of the DNA promoter protocadherin 8(PCDHg) gene in pancreatic cancer cell line Capan-2.The Capan-2 retardation in growth rate and apoptosis were assessed in when administered 5-Aza-CdR and the chemotherapy agent,gemcitabine.Methods MTT and flow cytometry were used to analyze the cell growth inhibition and apoptosis when treated with 5-Aza-CdR or in combination with gemcitabine.Methylation-specific PCR,RT-PCR and western blot were performed to detect methylation state,mRNA and protein respectively of PCDH8 gene in 5-Aza-CdR-treated Capan-2cells.Results Capan-2 cells treated with 5-Aza-CdR showed a slower growth rate,and a significant growth inhibition when given both 5-Aza-CdR in combination with gemcitabine.Compared with single drug administration and control,5-Aza-CdR together with gemcitabine can induce a stronger apoptosis signal.Different concentrations 5-Aza-CdR of were able to reverse methylation,restore mRNA and protein levels of PCDH8 in Capan-2.Conclusion 5 Aza-CdR may demethylate the PCDH8 gene,which would effectively remove the gene silencing caused by high methylation,and thus induce gene mRNA transcription and protein expression to inhibit cell growth and have collaborative antitumor functions with gemcitabine.
